Two exotic pests, Argentine stem weevil (ASW) and clover root weevil (CRW) are causing damage estimated at up to $200 M p.a. and $235 M p.a. respectively in dairy and sheep and beef pastures. While CRW is subject to successful biological control management it still causes considerable losses. Lesser pests also contribute to lost production, particularly as they often coexist with more major pests. However, their economic cost to New Zealand is difficult to calculate due to the variable nature of infestations on both temporal and spatial scales. At farm and paddock level, it is abundantly clear that substantial savings could be made if pest management is achieved. It is equally clear that in many instances the tools to do so are limited but if developed would contribute substantially to farm profitability.
The method whereby equal numbers of seven ecotypes of the parasitoid Microctonus hyperodae Loan (Hymenoptera: Braconidae, Euphorinae) were reared and released is described along with the reasons for doing so. This was achieved by variably intense rearing effort depending on the number of founder females in that particular ecotype.The parasitoid was released in three regions of New Zealand as a control agent of the Listronotus bonariensis Kuschel (Col. : Curculionidae), a severe pest of New Zealand pastures. It was later recovered from all three regions.
Purified RNA transcripts from venom glands dissected from the parasitoid wasp Microctonus hyperodae were copied, cloned and sequenced using traditional dideoxy sequencing methods. Using mass spectrometry analysis of the trypsinised PAGE gel protein bands we identified the RNA transcripts for the 3 most abundant proteins found in the venom and hence obtained their full protein sequence. Other abundant transcripts were also further sequenced. To reduce the effort required to obtain transcript information we dissected venom glands from a second parasitoid, Microctonus aethiopoides (Morocco biotype). The RNA transcripts were purified and reverse transcribed but instead of cloning the cDNA it was directly sequenced using Roche GS20 pyrosequencing. Results from a single GS20 sequencing run provided data similar to that obtained by the traditional methods used in analysing transcripts from M. hyperodae in a fraction of the time and cost. Comparing the transcripts between the two species showed that a similar range of genes are expressed with the putative orthologs of seven of the eight full length genes characterised from M. hyperodae being found in M. aethiopoides. Pyrosequencing should provide a valuable new method for rapidly sampling transcripts from a wide range of specialised insect tissues.
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